Analyzer for 2D Multiplex Detection of Infectious Agents

描述(由申请人提供)： 这笔赠款的目标是开发一种新的二维免疫遗传分析方法和芯片实验室设备，用于高度特异、敏感、定量和快速检测病原体。免疫遗传分析仪将用于受感染个体的定点诊断，用于食品、水和其他样本中病原体的分析点鉴定，并将开发一种用于高通量“临床”筛查的版本。我们的理由是，在所有这些应用中，至关重要的是要有一种具体的、假阳性很低的方法，这种方法可以量化污染的程度，并提供关于感染的基因信息，以便确定威胁的性质和治疗方法。这些长期目标将通过完成以下具体目标来实现：(1)将开发用于微流控捕获/预浓缩室的试剂、方法和微流控网络，这些微流控捕获/预浓缩室含有抗体装饰的珠子、整体或凝胶，用于捕获特定的细菌污染物(例如，来自我们的食品/水病原体开发小组)。样本将被泵过这些腔室，细菌将通过免疫捕获进行分离、浓缩和纯化。(2)这些微流控捕获方法将扩展到混合细菌种群的分离，使用一系列小室，每个小室针对不同类别的病原体。(3和4)将在面包板级别开发微流控方法，将免疫捕获小室与微型聚合酶链式反应扩增小室和毛细管电泳仪相结合，以提供感染的特异性、敏感性和定量，最重要的是提供感染的基因特征。(5)捕捉室、泵、聚合酶链式反应和毛细管电泳仪将作为便携式病原体分析仪的一部分整体集成在一个底物上，并将开发一系列仪器以提高性能和多重分析。(6)优化后的便携式系统将用于分析更多的致病细菌、病毒和细菌孢子。(7)此外，还将开发高通量、并行96个样本版本的免疫遗传分析仪。

DESCRIPTION (provided by applicant): The goal of this grant is to develop a novel two-dimensional immuno-genetic analysis method and lab-on-a-chip apparatus that will be used for highly specific, sensitive, quantitative and rapid detection of pathogens. The immuno-genetic analyzer will be used for point-of-care diagnosis of infected individuals, for point-of analysis identification of pathogens in food, water and other samples, and a version will be developed for high-throughput "clinical" screening. Our rationale is that in all these applications, it is critically important to have a method that is specific, that has very low false positives, that provides quantitation of the extent of contamination, and that provides genotypic information on the infection so that the nature of the threat as well as treatment are defined. These long-term goals will be approached by completing the following Specific Aims: (1) Reagents, methods and microfluidic networks will be developed for microfluidic capture/preconcentration chambers containing antibody decorated beads, monoliths or sol-gels that capture defined bacterial contaminants (e.g. from our development panel of food/water pathogens). Samples will be pumped through these chambers and the bacteria will be separated, concentrated and purified via immunocapture. (2) These microfluidic capture methods will be extended to the segregation of mixed bacterial populations by using a series of chambers each targeted to a different class of pathogens. (3 and 4) Microfluidic methods will be developed at the breadboard level for integrating immunocapture chambers with microfabricated PCR amplification chambers and capillary electrophoresis (CE) analyzers to provide specificity, sensitivity and quantitation and most importantly genetic characterization of the infection. (5) The capture chambers, pumps, PCR chambers and CE analyzers will be monolithically integrated on a substrate as a part of a portable pathogen analyzer, and a series of instruments will be developed to improve performance and multiplex analysis. (6) The optimized portable systems will be used to perform analyses of more pathogenic bacteria, viruses and bacterial spores. (7) In addition, a high-throughput, parallel 96 sample version of the immuno-genetic analyzer will be developed.